Multiplex real-time PCR assay for simultaneous quantification of BK polyomavirus, JC polyomavirus, and adenovirus DNA

Yasuhito Funahashi, Seiko Iwata, Yoshinori Ito, Seiji Kojima, Tetsushi Yoshikawa, Ryohei Hattori, Momokazu Gotoh, Yukihiro Nishiyama, Hiroshi Kimura

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

In recent years, virus-induced nephropathy caused mainly by BK polyomavirus (BKPyV), JC polyomavirus (JCPyV), and adenovirus has emerged as a problem in renal transplant patients. In the present study, we developed a multiplex real-time PCR assay to quantify the viral load of BKPyV, JCPyV, and adenovirus simultaneously. The dynamic range covered at least 6 orders of magnitude. This system was specific and reproducible, even in the presence of large amounts of DNA of other viruses. To validate this assay, urine samples from 124 renal transplant patients and serum samples from 18 hemorrhagic cystitis patients after hematopoietic stem cell transplantation were examined. In the urine samples from renal transplant patients, BKPyV was detected in 28 patients (22.6%), JCPyV was detected in 51 patients (41.1%), and adenovirus was detected in 2 patients (1.6%). The maximum amounts of each virus detected were 2.7 X 109, 8.7 X 108, and 1.2 X 102 copies/ml, respectively. Decoy cells were observed in 31 patients. The quantities of both BKPyV and JCPyV DNA were greater in samples with decoy cells. Two patients whose BKPyV viral loads exceeded 108 copies/ml had elevated serum creatinine levels and were diagnosed with BKPyV nephropathy based on graft biopsies. In serum samples from hemorrhagic cystitis patients, BKPyV, JCPyV, and adenovirus was detected in six, two, and three patients, respectively. Strong correlations were observed between the viral DNA copy numbers determined in the multiplex assays and those determined in single assays. Since this new assay is rapid, sensitive, and specific, it can be used for quantitative analyses of viruses in urine and serum samples after transplantation.

Original languageEnglish
Pages (from-to)825-830
Number of pages6
JournalJournal of Clinical Microbiology
Volume48
Issue number3
DOIs
Publication statusPublished - 01-03-2010

Fingerprint

BK Virus
JC Virus
Multiplex Polymerase Chain Reaction
Adenoviridae
Real-Time Polymerase Chain Reaction
DNA
Transplants
Cystitis
Urine
Viruses
Viral Load
Serum
Kidney
DNA Viruses
Hematopoietic Stem Cell Transplantation
Viral DNA
Creatinine

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

Cite this

Funahashi, Yasuhito ; Iwata, Seiko ; Ito, Yoshinori ; Kojima, Seiji ; Yoshikawa, Tetsushi ; Hattori, Ryohei ; Gotoh, Momokazu ; Nishiyama, Yukihiro ; Kimura, Hiroshi. / Multiplex real-time PCR assay for simultaneous quantification of BK polyomavirus, JC polyomavirus, and adenovirus DNA. In: Journal of Clinical Microbiology. 2010 ; Vol. 48, No. 3. pp. 825-830.
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Multiplex real-time PCR assay for simultaneous quantification of BK polyomavirus, JC polyomavirus, and adenovirus DNA. / Funahashi, Yasuhito; Iwata, Seiko; Ito, Yoshinori; Kojima, Seiji; Yoshikawa, Tetsushi; Hattori, Ryohei; Gotoh, Momokazu; Nishiyama, Yukihiro; Kimura, Hiroshi.

In: Journal of Clinical Microbiology, Vol. 48, No. 3, 01.03.2010, p. 825-830.

Research output: Contribution to journalArticle

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AU - Funahashi, Yasuhito

AU - Iwata, Seiko

AU - Ito, Yoshinori

AU - Kojima, Seiji

AU - Yoshikawa, Tetsushi

AU - Hattori, Ryohei

AU - Gotoh, Momokazu

AU - Nishiyama, Yukihiro

AU - Kimura, Hiroshi

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